Collection and Transportation of Microbiological samples

The steps of Collection and Transportation of Microbiological samples will be studied here.

Collection and transport of blood sample

  1. If using BacT/Alert bottles for bacteria or yeast, uncap the proper bottles, wipe the tops with alcohol and allow them to dry.
  2. If using Isolator tubes for unusual bacteria, fungi, or mycobacteria, disinfect the stopper with iodine and allow it to dry; do not let iodine pool in the stopper.
  3. Position the patient in a chair, or sitting or lying on a bed.
  4. Wash your hands.
  5. Apply a tourniquet to the patient’s extremity and locate a venipuncture site.
  6. Do not put the tourniquet on too tightly or leave it on the patient longer than 1 minute
  7. Next, put on non-latex gloves, and palpate for a vein.
  8. When a vein is selected, cleanse the area in a circular motion, beginning at the site and working outward. Allow the area to air dry. After the area is cleansed, it should not be touched or palpated again. If you find it necessary to reevaluate the site by palpation, the area needs to be re-cleansed before the venipuncture is performed
  9. Ask the patient to make a fist; avoid “pumping the fist.” Grasp the patient’s arm firmly using your thumb to draw the skin taut and anchor the vein. Swiftly insert the needle through the skin into the lumen of the vein. The needle should form a 15-30 degree angle with the arm surface.
  10. Remove enough blood to properly inoculate the bottle or tube: 8-10 ml per bottle/tube for adults, 0.5-4.0 ml for children.
  11. Remove the tourniquet, then the needle, and apply pressure to the venipuncture site.
  12. Cover with a pressure bandage and band-aid (Use surgical tape or paper tape if the patient is allergic to band-aid adhesive).
    Inoculate the bottles/tubes; the anaerobe bottle should be filled first, avoiding the introduction of air.
  13. Gently mix the bottles/tubes by inversion.
  14. Label the bottles/tubes and properly complete the request form noting the date, exact venipuncture time, and site.
  15. Place in plastic bag and transport to the Microbiology Laboratory within 30 minutes.

Collection and transport of Sputum sample

 An early morning specimen is recommended. A volume of 5 to 10 ml is adequate and there is no advantage in collecting a larger volume. The sample should contain recently discharged material from the bronchial tree with minimal saliva content.

  1. Upon rising in the morning, rinse the mouth well with water (not the mouthwash).
  2. Inhale as deeply as possible. Expectorate into a sputum collection container, available from the Laboratory, while coughing as deeply and vigorously as possible.
  3. Tightly cap container so no spillage occurs.
  4. Wash and dry outside of the container and label the SPECIMEN CONTAINER with name and date and time of collection.
  5. Complete the requisition form
  6. Specimens must be processed by the laboratory within 24 hours.
  7. If more than one specimen is requested (for example, 3 AFB cultures are requested), collect only one specimen per day, first thing in the morning. Have specimens delivered to the Laboratory DAILY. Do not wait until all three have been collected.

Collection and transport of Nasopharyngeal sample

  1. Collect specimen under good lighting. Bend the wire of the nasopharyngeal swab into a semi-circle and pass through the nostril to the pharynx, scrub back and forth gently 2 3 times and remove.
  2. Return the swab to the transport tube (follow the manufacturer’s directions).
  3. Label the swab transport tube with a unique patient identifier.
  4. Complete the requisition form.
  5. Place specimen and requisition properly and arrange to have specimen taken to a courier pickup site. DO NOT mail specimen.

Collection and transport of THROAT SWAB

  1. Collect specimen under good lighting. Depress the tongue with a tongue blade and pass the swab firmly over the back of the patient’s throat, tonsils, and any area of inflammation and or exudation.
  2. Return the swab to the transport tube
  3. Label the swab transport tube with the patient name.
  4. Complete the requisition form or order test directly in LIMS.
  5. Place specimen and requisition properly and arrange to have specimen taken to a courier pickup site. DO NOT mail specimen.

(for 8hrs before swabbing, patient should not be treated with antibiotics or antiseptic mouth-washes; organisms may be temporarily absent)

Collection and transport of WOUND (Pus) samples

  1. Collect specimen under good lighting. Pass the swab firmly over or into an area of suspected infection and obtain a sample of exudate, drainage, or purulent discharge if these are present.
  2. Return the swab to the transport tube
  3. Label the swab transport tube with a unique patient identifier.
  4. Complete the requisition form. Indicate the area of the body from which wound specimen was taken to assist in distinguishing normal from abnormal flora (ears, legs, etc).
  5. Place specimen and requisition in the pouch and promptly deliver to the lab pickup site. DO NOT mail specimen.
    • NOTE: Specimens collected on culturettes should be delivered to the laboratory within 24 hours in order that they are processed immediately after collection. Culturettes 24-72 hours old are acceptable if refrigerated.

Collection and Transport of TISSUE sample

Handle in the same manner as wound specimens above; however, instead of culturette (sterile swab tube), place tissue specimen in sterile container with a small amount of sterile saline or sterile water to keep specimen from drying out. Be sure container is labeled with name of patient and source of specimen.

Collection and Transport of STOOL sample

The recovery of bacterial pathogens from fecal specimens will help confirm the diagnosis of bacterial gastroenteritis as manifested by diarrhea and/or dysentery. Stools are routinely screened for Campylobacter, Salmonella, Shigella, and Shiga toxin E.coli. If other pathogens are suspected indicate on the request form.

 For Norovirus stool collection – collect teaspoon size sample in sterile leak-proof container with no preservatives along with test requisition form. Store samples in refrigerator at (3-9)C

1. give the patient a clean, dry, disinfectant-free suitable wide-necked container to pass the feces or a kitchen wrap can be used on a toilet pan for collection; ask them not to contaminate with urine

2. transfer a portion of (about  a spoonful for formed and no more than 10ml for liquid stool) of specimen (specifically containing mucus, pus or blood) into a clean, dry leak-proof  container containing transport medium (Cary – Blair transport medium); mix thoroughly

(in case of worms, transfer the specimen with worm segments or eggs in a separate container and send to lab for identification)

3. on the requisition form, write the color, and whether the specimen is formed, semi-formed, unformed or fluid (liquid); also report if blood, mucus or worms segments are present.

Collection and Transport of URINE sample

Whenever possible, the first urine passed by patient at the beginning of the day should be sent to lab as it is highly concentrated and thus most suitable for culture, microscopy and biochemical analysis.

1. a sterile, dry, wide-necked, leak proof container is provided to the patient and requested for (10-20)ml  Mid- Stream Urine

2. Wash hands with soap and water, rinse and dry.

3. WASH area around urethra with soap.

4. RINSE area with warm water.

5. VOID- Pass the first portion of urine into the toilet and then pass a portion (15-20ml) of the remaining urine into a sterile container. Pass the rest of the urine into the toilet, close and label the container with name and date and time of collection. Deliver immediately to the laboratory  or if delay is anticipated, store samples in refrigerator (4-6)C.

Collection and Transport of GENITAL TRACT specimens

  1. Use swab to obtain a sample of endocervical, vaginal, or urethral discharge. Return the swab to the transport tube and break the media ampule at the base of the tube to moisten the swab. Please follow the manufacturer’s directions.
  2. Label the swab transport tube with the patient’s name. Complete the requisition form
  3. Place specimen and requisition in pouch and transport to the lab DO NOT mail specimen.
  4. Special media and transport containers are required when culturing for gonorrhea.

Collection and Transport of CEREBROSPINAL FLUID

Must be collected by an experienced medical officer or health worker; collected aseptically to prevent organisms into central nervous system.

1. fluid is collected from aranchoid space; a sterile, wide- bore needle is inserted between 4th and 5th lumbar vertebrae and CSF is allowed to drip into a dry sterile container

2. two sterile, screw-capped containers are used, one for culture and next for other tests; 1ml CSF in culture  tube and (2-3)ml in other

3. Immediate processing is carried out in laboratory

* Things to remember

  • Specimens for microbiological investigations should be delivered to laboratory without delay and processed as soon as possible (prevent contamination)
  • If delay is unavoidable, suitable transport medium must be used; helps organism from dying due to enzyme action, change in pH or lack of essential nutrients.
  • Specimens/samples must be packed well and safely before transport; keep a register of all specimens dispatched, the name, number, address of patient, type of specimen, investigation required, date of dispatch
  • Check the specimen container is free from cracks, and the cap is leak-proof, seal around the container cap with adhesive tape to prevent leakage during transit; mark all the specimen that may contain highly infectious organism as ‘HIGH RISK’

Binod G C

I'm Binod G C (MSc), a PhD candidate in cell and molecular biology who works as a biology educator and enjoys scientific blogging. My proclivity for blogging is intended to make notes and study materials more accessible to students.

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